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Abstract
During the last few years, an important element in the improvement of the molecular biology techniques has been the necessity for availability of high quality and functionality DNA. Several DNA extraction procedures with different results in both performance and quality, have been proposed. In this study our objective was to determine the most reliable extraction method that balances DNA quantity, and to assess the sample quantification of the fluorometric DNA quantification methods. For this, blood extracted by venopunction from 20 healthy volunteers was used to obtain DNA from buffy coat, and 4 commercial DNA extraction kits were assessed as well as two fluorometric DNA quantification methods with protocols of different complexity. Results suggest that manual methods achieve higher quality and larger yields of DNA. DNA purity obtained with the 4 extraction kits evaluated through the 260/280 and 260/230 ratio showed that the Qiacube kit fulfilled the criteria established in this work, followed very close by the Flexigene kit. On the other hand, the fluorometric DNA methods used in the samples quantification showed a higher variability when using QuantiFlour method, obtaining better results probably due to the simplicity of this protocol.
Acknowledgments
The authors thank all technical personnel of the genetic platform of the Instituto de Investigación Biomédica de Málaga (IBIMA).
Informed consent
All individuals gave written informed consent to participate in research prior to blood collection in accordance with the Andalusian Public Health System Biobank located in Málaga, and approved by the Ethics Committee of Clinical Research (CEIC) from IBIMA.
Disclosure statement
No potential conflict of interest was reported by the author(s).